Medium and large-sized terrestrial mammals develop important functions in ecosystems maintenance. However, this group is the most vulnerable to local extinctions, through habitat loss or hunting. This study inventoried the community of medium and large mammals in the Reserva Ecológica de Guapiaçú (REGUA), Cachoeiras de Macacu, RJ, providing data on species composition, richness and abundance. Species were registered through the use of camera traps, sign survey and visual encounters on pre-existing trails and roads. The study was conducted from January to October 2012, with each month considered as a sample unit. An effort of 1568 cameras-day and 120 km traveled, resulted on 302 records of 22 species of medium and large-sized mammals, belonging to eight orders. This value corresponds to more than three-quarters of the richness described from this group in the Atlantic Forest of Rio de Janeiro state. Five species are under some level of regional, national or global threat. Three game species, Pecari tajacu, Cuniculus paca and Dasyprocta leporina were abundant in the area. The observed richness was comparable to that found in other studies conducted in rain forests. Thus, for its high number of species, including those under some degree of threat, we conclude that REGUA is an important area to mammal conservation, especially in the Atlantic Forest of Rio de Janeiro state.